Classifications

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  • C07D—HETEROCYCLIC COMPOUNDS
  • C07D491/00—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00
  • C07D491/02—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00 in which the condensed system contains two hetero rings
  • C07D491/04—Ortho-condensed systems
  • C07D491/044—Ortho-condensed systems with only one oxygen atom as ring hetero atom in the oxygen-containing ring
  • C07D491/048—Ortho-condensed systems with only one oxygen atom as ring hetero atom in the oxygen-containing ring the oxygen-containing ring being five-membered
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  • C07D—HETEROCYCLIC COMPOUNDS
  • C07D491/00—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00
  • C07D491/02—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00 in which the condensed system contains two hetero rings
  • C07D491/06—Peri-condensed systems
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  • C07—ORGANIC CHEMISTRY
  • C07D—HETEROCYCLIC COMPOUNDS
  • C07D491/00—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00
  • C07D491/02—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00 in which the condensed system contains two hetero rings
  • C07D491/04—Ortho-condensed systems
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K31/00—Medicinal preparations containing organic active ingredients
  • A61K31/33—Heterocyclic compounds
  • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
  • A61K31/535—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with at least one nitrogen and one oxygen as the ring hetero atoms, e.g. 1,2-oxazines
  • A61K31/5375—1,4-Oxazines, e.g. morpholine
  • A61K31/5377—1,4-Oxazines, e.g. morpholine not condensed and containing further heterocyclic rings, e.g. timolol
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P35/00—Antineoplastic agents
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07D—HETEROCYCLIC COMPOUNDS
  • C07D413/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms
  • C07D413/14—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing three or more hetero rings
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07D—HETEROCYCLIC COMPOUNDS
  • C07D471/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00
  • C07D471/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00 in which the condensed system contains two hetero rings
  • C07D471/04—Ortho-condensed systems
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07D—HETEROCYCLIC COMPOUNDS
  • C07D471/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00
  • C07D471/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00 in which the condensed system contains two hetero rings
  • C07D471/06—Peri-condensed systems
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07D—HETEROCYCLIC COMPOUNDS
  • C07D471/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00
  • C07D471/12—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00 in which the condensed system contains three hetero rings
  • C07D471/14—Ortho-condensed systems
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07D—HETEROCYCLIC COMPOUNDS
  • C07D471/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00
  • C07D471/12—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00 in which the condensed system contains three hetero rings
  • C07D471/16—Peri-condensed systems
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07D—HETEROCYCLIC COMPOUNDS
  • C07D487/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00
  • C07D487/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00 in which the condensed system contains two hetero rings
  • C07D487/04—Ortho-condensed systems
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07D—HETEROCYCLIC COMPOUNDS
  • C07D487/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00
  • C07D487/12—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00 in which the condensed system contains three hetero rings
  • C07D487/14—Ortho-condensed systems
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07D—HETEROCYCLIC COMPOUNDS
  • C07D491/00—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00
  • C07D491/12—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00 in which the condensed system contains three hetero rings
  • C07D491/14—Ortho-condensed systems
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07D—HETEROCYCLIC COMPOUNDS
  • C07D491/00—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00
  • C07D491/12—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00 in which the condensed system contains three hetero rings
  • C07D491/14—Ortho-condensed systems
  • C07D491/147—Ortho-condensed systems the condensed system containing one ring with oxygen as ring hetero atom and two rings with nitrogen as ring hetero atom
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07D—HETEROCYCLIC COMPOUNDS
  • C07D491/00—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00
  • C07D491/12—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00 in which the condensed system contains three hetero rings
  • C07D491/16—Peri-condensed systems

Definitions

• Epigenetic regulation of gene expression is an important biological determinant of protein production and cellular differentiation and plays a significant pathogenic role in a number of human diseases.
• Epigenetic regulation involves heritable modification of genetic material without changing its nucleotide sequence.
• epigenetic regulation is mediated by selective and reversible modification (e.g., methylation) of DNA and proteins (e.g., histones) that control the conformational transition between transcriptionally active and inactive states of chromatin.
• methyltransferases e.g., PRMT5
• PRMT5 plays a role in diseases such as proliferative disorders, metabolic disorders, and blood disorders.
• the homozygous deletion of tumor suppressor genes is a key driver of cancer, frequently resulting in the collateral loss of passenger genes located in close genomic proximity to the tumor suppressor. Deletion of these passenger genes can create therapeutically tractable vulnerabilities that are specific to tumor cells. Homozygous deletion of the chromosome 9p21 locus, which harbors the well-known tumor suppressor CDKN2A (cyclin dependent kinase inhibitor 2A), occurs in 15% of all tumors and frequently includes the passenger gene MTAP (methylthioadenosine phosphorylase), a key enzyme in the methionine and adenine salvage pathways. Deletion of MTAP results in accumulation of its substrate, methylthioadenosine (MTA).
• MTA methylthioadenosine
• MTA shares close structural similarity to S-adenosylmethionine (SAM), the substrate methyl donor for the type II methyltransferase PRMT5. Elevated MTA levels, driven by loss of MTAP, selectively compete with SAM for binding to PRMT5, placing the methyltransferase in a hypomorphic state, vulnerable to further PRMT5 inhibition.
• SAM S-adenosylmethionine
• Multiple genome scale shRNA drop out screens performed in large tumor cell line panels have identified a strong correlation between MTAP loss and cell line dependency on PRMT5, further highlighting the strength of this metabolic vulnerability.
• PRMT5 is a known cell essential gene and conditional PRMT5 knockout and siRNA knockdown studies suggest that significant liabilities could be associated with inhibiting PRMT5 in normal tissues (e.g., pan-cytopenia, infertility, skeletal muscle loss, cardiac hypertrophy). Therefore, novel strategies are required to exploit this metabolic vulnerability and preferentially target PRMT5 in MTAP null tumors while sparing PRMT5 in normal tissues (MTAP WT).
• Targeting PRMT5 with an MTA-cooperative small molecule inhibitor could preferentially target the MTA bound state of PRMT5, enriched in MTAP null tumor cells, while providing an improved therapeutic index over normal cells where MTAP is intact and MTA levels are low.
• the invention provides a compound of Formula I
• the invention provides compounds, the tautomer thereof, the stereoisomer thereof, or the pharmaceutically acceptable salt of any of the foregoing, wherein R is a tricycle of Formula IA
• R can be any organic radical
• R can be
• R can be any organic compound
• the invention provides compounds, the tautomer thereof, the stereoisomer thereof, or the pharmaceutically acceptable salt of any of the foregoing, wherein R is
• R can be any organic radical
• R can be any organic compound
• the invention encompasses compound as described above, the tautomer thereof, the stereoisomer thereof, or the pharmaceutically acceptable salt of any of the foregoing, wherein X 1 is C.
• X 1 can be substituted with halo.
• the invention further encompasses compound as described above, the tautomer thereof, the stereoisomer thereof, or the pharmaceutically acceptable salt of any of the foregoing, wherein X 1 can be N.
• the invention further provides compounds, the tautomer thereof, the stereoisomer thereof, or the pharmaceutically acceptable salt of any of the foregoing, wherein R is a tricycle of Formula IB
• X 7 can be N.
• R can be any organic compound
• R can be any organic compound
• the invention further provides compounds as described above, the tautomer thereof, the stereoisomer thereof, or the pharmaceutically acceptable salt of any of the foregoing, wherein R 1 , R 2 and the nitrogen atom to which they are attached form a six membered ring that may be saturated or partially saturated, and comprises 0, 1 or 2 additional heteroatoms independently selected from O, N or S, wherein the six membered ring is
• R 3 can be selected from phenyl, pyridinyl, pyrazidinyl, or pyrimidinyl, optionally independently substituted with one or more R.
• R 3 can be phenyl, optionally independently substituted with one or more R a .
• R a can be, for example, C 1-6 haloalkyl or —OC 1-3 haloalkyl.
• halo can be Cl or F.
• the invention provides compounds the tautomer thereof, the stereoisomer thereof, or the pharmaceutically acceptable salt of any of the foregoing, wherein R 1 , R 2 and the nitrogen atom to which they are attached form a six membered ring that may be saturated or partially saturated, and comprises 0, 1 or 2 additional heteroatoms selected from O, N or S, wherein the six membered ring can be
• the six membered ring can be
• R 3 can be independently selected from phenyl, pyridinyl, pyrazidinyl or pyrimidinyl, optionally independently substituted with one or more R a .
• the invention provides compounds, the tautomer thereof, the stereoisomer thereof, or the pharmaceutically acceptable salt of any of the foregoing, wherein R 1 , R 2 and the nitrogen atom to which they are attached form pyrrolidinyl.
• these compounds include R 3 as phenyl, optionally independently substituted with one or more R a .
• R a can be C 1-6 haloalkyl or —OC 1-3 haloalkyl.
• the invention encompasses compounds as described above, the tautomer thereof, the stereoisomer thereof, or the pharmaceutically acceptable salt of any of the foregoing, wherein X 1 is C, optionally substituted with C 1-3 alkyl or halo.
• C 1-3 alkyl can be methyl.
• halo can be Cl or Br.
• the invention encompasses compounds as described above, the tautomer thereof, the stereoisomer thereof, or the pharmaceutically acceptable salt of any of the foregoing, wherein X 1 is N.
• the invention provides compounds as described above, the tautomer thereof, the stereoisomer thereof, or the pharmaceutically acceptable salt of any of the foregoing, wherein R 1 , R 2 and the nitrogen atom to which they are attached form a six membered ring that may be saturated or partially saturated, and comprises 0, 1 or 2 additional heteroatoms selected from O, N or S, wherein the six membered ring can be
• R 3 can be independently selected from phenyl, pyridinyl, pyrazidinyl and pyrimidinyl, optionally substituted with one or more R a .
• R a can be in each instance selected from halo, CN, C 1-6 alkyl, C 1-6 haloalkyl, —OC 1-3 alkyl, and —OC 1-3 haloalkyl.
• the invention provides compounds as described above, the tautomer thereof, the stereoisomer thereof, or the pharmaceutically acceptable salt of any of the foregoing, wherein R 1 , R 2 and the nitrogen atom to which they are attached form a six membered ring that may be saturated or partially saturated, and comprises 0, 1 or 2 additional heteroatoms selected from O, N or S, wherein the six membered ring can be
• R 3 can be independently selected from phenyl, pyridinyl, pyrazidinyl and pyrimidinyl, optionally substituted with one or more R a .
• R a can be in each instance independently selected from halo, CN, C 1-6 alkyl, C 1-6 haloalkyl, —OC 1-3 alkyl, and —OC 1-3 haloalkyl.
• R 3 can be pyridinyl, optionally substituted with one or more R a .
• R a can be C 1-6 haloalkyl.
• R a can be pentafluorosulfanyl.
• the invention provides the compound, the tautomer thereof, the stereoisomer thereof, or the pharmaceutically acceptable salt of any of the foregoing, wherein the compound is selected from:
• the invention further provides methods of treating cancer comprising administering to a subject an effective amount of the compound of the invention, the tautomer thereof, the stereoisomer thereof, or the pharmaceutically acceptable salt of any of the foregoing.
• the cancer is selected from lung, Head and Neck Squamous Cell Carcinoma (HNSCC), esophagus, lymphoid, glioblastoma, colon, melanoma, gastric, pancreatic, bile or bladder cancer.
• lung cancer could be Non-Small Cell Lung Carcinoma (NSCLC).
• the invention further provides pharmaceutical compositions, comprising the compounds of the invention, the tautomer thereof, the stereoisomer thereof, or the pharmaceutically acceptable salt of any of the foregoing or a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable excipient.
• the invention also provides methods of manufacturing a medication for treating a cancer, the method comprising administering to a subject an effective amount of the compound of the invention, the tautomer thereof, the stereoisomer thereof, or the pharmaceutically acceptable salt of any of the foregoing.
• the cancer can be lung, Head and Neck Squamous Cell Carcinoma (HNSCC), esophagus, lymphoid, glioblastoma, colon, bile, melanoma, gastric, pancreatic or bladder cancer.
• lung cancer could be Non-Small Cell Lung Carcinoma (NSCLC).
• the invention also provides the compound of the invention, the tautomer thereof, the stereoisomer thereof, or the pharmaceutically acceptable salt of any of the foregoing for use in a method of treating a cancer, the method comprising administering to a subject an effective amount of such compound.
• the cancer can lung, Head and Neck Squamous Cell Carcinoma (HNSCC), esophagus, lymphoid, glioblastoma, colon, melanoma, gastric, pancreatic bile or bladder cancer.
• lung cancer could be Non-Small Cell Lung Carcinoma (NSCLC).
• the cancer can be lung, Head and Neck Squamous Cell Carcinoma (HNSCC), esophagus, lymphoid, glioblastoma, colon, melanoma, gastric, pancreatic, bile or bladder cancer.
• lung cancer could be Non-Small Cell Lung Carcinoma (NSCLC).
• any variable occurs more than one time in a chemical formula, its definition on each occurrence is independent of its definition at every other occurrence. If the chemical structure and chemical name conflict, the chemical structure is determinative of the identity of the compound.
• the compounds of the present disclosure may contain one or more chiral centers and/or double bonds and therefore, may exist as stereoisomers, such as double-bond isomers (i.e., geometric isomers), enantiomers, or diastereomers.
• any chemical structures within the scope of the specification depicted, in whole or in part, with a relative configuration encompass all possible enantiomers and stereoisomers of the illustrated compounds including the stereoisomerically pure form (e.g., geometrically pure, enantiomerically pure or diastereomerically pure) and enantiomeric and stereoisomeric mixtures.
• Enantiomeric and stereoisomeric mixtures can be resolved into the component enantiomers or stereoisomers using separation techniques or chiral synthesis techniques well known to the skilled artisan.
• Certain compounds of the invention may possess asymmetric carbon atoms (optical centers) or double bonds; the racemates, enantiomers, diastereomers, geometric isomers and individual isomers are all intended to be encompassed within the scope of the invention.
• atropisomers and mixtures thereof such as those resulting from restricted rotation about two aromatic or heteroaromatic rings bonded to one another are intended to be encompassed within the scope of the invention.
• substituent is a phenyl group and is substituted with two groups bonded to the C atoms adjacent to the point of attachment to the N atom of the triazole, then rotation of the phenyl may be restricted.
• the barrier of rotation is high enough that the different atropisomers may be separated and isolated.
• stereoisomer or “stereomerically pure” means one stereoisomer of a compound that is substantially free of other stereoisomers of that compound.
• a stereomerically pure compound having one chiral center will be substantially free of the mirror image enantiomer of the compound.
• a stereomerically pure compound having two chiral centers will be substantially free of other diastereomers of the compound.
• a typical stereomerically pure compound comprises greater than about 80% by weight of one stereoisomer of the compound and less than about 20% by weight of other stereoisomers of the compound, more preferably greater than about 90% by weight of one stereoisomer of the compound and less than about 10% by weight of the other stereoisomers of the compound, even more preferably greater than about 95% by weight of one stereoisomer of the compound and less than about 5% by weight of the other stereoisomers of the compound, and most preferably greater than about 97% by weight of one stereoisomer of the compound and less than about 3% by weight of the other stereoisomers of the compound.
• stereochemistry of a structure or a portion of a structure is not indicated with, for example, bold or dashed lines, the structure or portion of the structure is to be interpreted as encompassing all stereoisomers of it.
• a bond drawn with a wavy line indicates that both stereoisomers are encompassed. This is not to be confused with a wavy line drawn perpendicular to a bond which indicates the point of attachment of a group to the rest of the molecule.
• certain compounds of the invention may exist in one or more tautomeric forms. Because one chemical structure may only be used to represent one tautomeric form, it will be understood that for convenience, referral to a compound of a given structural formula includes tautomers of the structure represented by the structural formula. Depending on the compound, some compounds may exist primarily in one form more than another. Also, depending on the compound and the energy required to convert one tautomer to the other, some compounds may exist as mixtures at room temperature whereas others may be isolated in one tautomeric form or the other.
• tautomers associated with compounds of the invention are those with a pyridone group (a pyridinyl) for which hydroxypyridine is a tautomer and compounds with a ketone group with the enol tautomer. Examples of these are shown below.
• Compounds of the present disclosure include, but are not limited to, compounds of Formula I and all pharmaceutically acceptable forms thereof.
• the invention includes pharmaceutically acceptable forms of the compounds pharmaceutically acceptable salts, solvates, crystal forms (including polymorphs and clathrates), chelates, non-covalent complexes, prodrugs, and mixtures thereof.
• the invention discloses compounds, and a pharmaceutically acceptable salt thereof, a solvate thereof, a chelate thereof, a non-covalent complex thereof, a prodrug thereof, and ester prodrugs such as (C 1 -C 4 )alkyl esters and mixtures of any of the foregoing.
• Pharmaceutically acceptable salts of the compounds of the present invention include acid addition salts formed with inorganic acids such as hydrochloric, hydrobromic, hydroiodic, phosphoric, metaphosphoric, nitric and sulfuric acids, and with organic acids, such as tartaric, acetic, trifluoroacetic, citric, malic, lactic, fumaric, benzoic, formic, propionic, glycolic, gluconic, maleic, succinic, camphorsulfuric, isothionic, mucic, gentisic, isonicotinic, saccharic, glucuronic, furoic, glutamic, ascorbic, anthranilic, salicylic, phenylacetic, mandelic, embonic (pamoic), methanesulfonic, ethanesulfonic, pantothenic, stearic, sulfinilic, alginic, galacturonic and arylsulfonic, for example benzenesulfonic
• Suitable salts include those described in P. Heinrich Stahl, Camille G. Wermuth (Eds.), Handbook of Pharmaceutical Salts Properties, Selection and Use; 2002. Salts having a non-pharmaceutically acceptable anion or cation are within the scope of the invention as useful intermediates for the preparation of pharmaceutically acceptable salts and/or for use in non-therapeutic, for example, in vitro, situations.
• solvate refers to the compound formed by the interaction of a solvent and a compound. Solvates of a compound includes solvates of all forms of the compound. In certain embodiments, solvents are volatile, non-toxic, and/or acceptable for administration to humans in trace amounts. Suitable solvates are pharmaceutically acceptable solvates, such as hydrates, including monohydrates and hemi-hydrates.
• the invention discloses compounds which may also contain naturally occurring or unnatural proportions of atomic isotopes at one or more of the atoms that constitute such compounds.
• the compounds may be radiolabeled with radioactive isotopes, such as for example tritium (H), iodine-125 ( 125 I) or carbon-14 ( 14 C).
• Radiolabeled compounds are useful as therapeutic or prophylactic agents, research reagents, e.g., assay reagents, and diagnostic agents, e.g., in vivo imaging agents. All isotopic variations of the compounds of the invention, whether radioactive or not, are intended to be encompassed within the scope of the invention.
• the invention also includes deuterium (D) or tritium (T) containing compounds.
• Alkyl refers to a saturated branched or straight-chain monovalent hydrocarbon group derived by the removal of one hydrogen atom from a single carbon atom of a parent alkane.
• Typical alkyl groups include, but are not limited to, methyl, ethyl, propyls such as propan-1-yl and propan-2-yl, butyls such as butan-1-yl, butan-2-yl, 2-methyl-propan-1-yl, 2-methyl-propan-2-yl, tert-butyl, and the like.
• an alkyl group comprises 1 to 20 carbon atoms.
• alkyl groups include 1 to 10 carbon atoms or 1 to 6 carbon atoms whereas in other embodiments, alkyl groups include 1 to 4 carbon atoms. In still other embodiments, an alkyl group includes 1 or 2 carbon atoms. Branched chain alkyl groups include at least 3 carbon atoms and typically include 3 to 7, or in some embodiments, 3 to 6 carbon atoms. An alkyl group having 1 to 6 carbon atoms may be referred to as a (C 1 -C 6 )alkyl group and an alkyl group having 1 to 4 carbon atoms may be referred to as a (C 1 -C 4 )alkyl. This nomenclature may also be used for alkyl groups with differing numbers of carbon atoms.
• Alkenyl refers to an unsaturated branched or straight-chain hydrocarbon group having at least one carbon-carbon double bond derived by the removal of one hydrogen atom from a single carbon atom of a parent alkene.
• the group may be in either the Z- or E-form (cis or trans) about the double bond(s).
• Typical alkenyl groups include, but are not limited to, ethenyl; propenyls such as prop-1-en-1-yl, prop-1-en-2-yl, prop-2-en-1-yl (allyl), and prop-2-en-2-yl; butenyls such as but-1-en-1-yl, but-1-en-2-yl, 2-methyl-prop-1-en-1-yl, but-2-en-1-yl, but-2-en-1-yl, but-2-en-2-yl, buta-1,3-dien-1-yl, and buta-1,3-dien-2-yl; and the like.
• an alkenyl group has 2 to 20 carbon atoms and in other embodiments, has 2 to 6 carbon atoms.
• An alkenyl group having 2 to 6 carbon atoms may be referred to as a (C 2 -C 6 )alkenyl group.
• Alkynyl refers to an unsaturated branched or straight-chain hydrocarbon having at least one carbon-carbon triple bond derived by the removal of one hydrogen atom from a single carbon atom of a parent alkyne.
• Typical alkynyl groups include, but are not limited to, ethynyl; propynyl; butynyl, 2-pentynyl, 3-pentynyl, 2-hexynyl, 3-hexynyl and the like.
• an alkynyl group has 2 to 20 carbon atoms and in other embodiments, has 2 to 6 carbon atoms.
• An alkynyl group having 2 to 6 carbon atoms may be referred to as a —(C 2 -C 6 )alkynyl group.
• Alkoxy refers to a radical —OR where R represents an alkyl group as defined herein. Representative examples include, but are not limited to, methoxy, ethoxy, propoxy, butoxy, and the like. Typical alkoxy groups include 1 to 10 carbon atoms, 1 to 6 carbon atoms or 1 to 4 carbon atoms in the R group. Alkoxy groups that include 1 to 6 carbon atoms may be designated as —O—(C 1 -C 6 ) alkyl or as —O—(C 1 -C 6 alkyl) groups. In some embodiments, an alkoxy group may include 1 to 4 carbon atoms and may be designated as —O—(C 1 -C 4 ) alkyl or as —O—(C 1 -C 4 alkyl) groups group.
• Aryl refers to a monovalent aromatic hydrocarbon group derived by the removal of one hydrogen atom from a single carbon atom of a parent aromatic ring system.
• Aryl encompasses monocyclic carbocyclic aromatic rings, for example, benzene.
• Aryl also encompasses bicyclic carbocyclic aromatic ring systems where each of the rings is aromatic, for example, naphthalene.
• Aryl groups may thus include fused ring systems where each ring is a carbocyclic aromatic ring.
• an aryl group includes 6 to 10 carbon atoms. Such groups may be referred to as C 6 -C 10 aryl groups.
• Aryl does not encompass or overlap in any way with heteroaryl as separately defined below. Hence, if one or more carbocyclic aromatic rings is fused with an aromatic ring that includes at least one heteroatom, the resulting ring system is a heteroaryl group, not an aryl group, as defined herein.
• Carbonyl refers to the radical —C(O) which may also be referred to as —C( ⁇ O) group.
• Carboxy refers to the radical —C(O)OH which may also be referred to as —C( ⁇ O)OH.
• “Cyano” refers to the radical —CN.
• Cycloalkyl refers to a saturated cyclic alkyl group derived by the removal of one hydrogen atom from a single carbon atom of a parent cycloalkane.
• Typical cycloalkyl groups include, but are not limited to, groups derived from cyclopropane, cyclobutane, cyclopentane, cyclohexane, cycloheptane, cyclooctane, and the like. Cycloalkyl groups may be described by the number of carbon atoms in the ring.
• a cycloalkyl group having 3 to 8 ring members may be referred to as a (C 3 -C 8 )cycloalkyl
• a cycloalkyl group having 3 to 7 ring members may be referred to as a (C 3 -C 7 )cycloalkyl
• a cycloalkyl group having 4 to 7 ring members may be referred to as a (C 4 -C 7 )cycloalkyl.
• the cycloalkyl group can be a (C 3 -C 10 )cycloalkyl, a (C 3 -C 8 )cycloalkyl, a (C 3 -C 7 )cycloalkyl, a (C 3 -C 6 )cycloalkyl, or a (C 4 -C 7 )cycloalkyl group and these may be referred to as C 3 -C 10 cycloalkyl, C 3 -C 8 cycloalkyl, C 3 -C 7 cycloalkyl, C 3 -C 6 cycloalkyl, or C 4 -C 7 cycloalkyl groups using alternative language.
• Heterocyclyl refers to a cyclic group that includes at least one saturated, partially unsaturated, cyclic ring. Heterocyclyl groups include at least one heteroatom as a ring member. Typical heteroatoms include O, S and N and are independently chosen. Heterocyclyl groups include monocyclic ring systems and bicyclic ring systems. Bicyclic heterocyclyl groups include at least one non-aromatic ring with at least one heteroatom ring member that may be fused to a cycloalkyl ring or may be fused to an aromatic ring where the aromatic ring may be carbocyclic or may include one or more heteroatoms.
• the point of attachment of a bicyclic heterocyclyl group may be at the non-aromatic cyclic ring that includes at least one heteroatom or at another ring of the heterocyclyl group.
• a heterocyclyl group derived by removal of a hydrogen atom from one of the 9 membered heterocyclic compounds shown below may be attached to the rest of the molecule at the 5-membered ring or at the 6-membered ring.
• a heterocyclyl group includes 5 to 10 ring members of which 1, 2, 3 or 4 are heteroatoms independently selected from O, S, or N. In other embodiments, a heterocyclyl group includes 3 to 7 ring members of which 1, 2, or 3 heteroatom are independently selected from O, S, or N. In such 3-7 membered heterocyclyl groups, only 1 of the ring atoms is a heteroatom when the ring includes only 3 members and includes 1 or 2 heteroatoms when the ring includes 4 members. In some embodiments, a heterocyclyl group includes 3 or 4 ring members of which 1 is a heteroatom selected from O, S, or N.
• a heterocyclyl group includes 5 to 7 ring members of which 1, 2, or 3 are heteroatoms independently selected from O, S, or N.
• Typical heterocyclyl groups include, but are not limited to, groups derived from epoxides, aziridine, azetidine, imidazolidine, morpholine, piperazine, piperidine, hexahydropyrimidine, 1,4,5,6-tetrahydropyrimidine, pyrazolidine, pyrrolidine, quinuclidine, tetrahydrofuran, tetrahydropyran, benzimidazolone, pyridinone, and the like.
• Heterocyclyl groups may be fully saturated but may also include one or more double bonds.
• heterocyclyl groups include, but are not limited to, 1,2,3,6-tetrahydropyridinyl, 3,6-dihydro-2H-pyranyl, 3,4-dihydro-2H-pyranyl, 2,5-dihydro-1H-pyrolyl, 2,3-dihydro-1H-pyrolyl, 1H-azirinyl, 1,2-dihydroazetenyl, and the like.
• Substituted heterocyclyl also includes ring systems substituted with one or more oxo ( ⁇ O) or oxide (—O—) substituents, such as piperidinyl N-oxide, morpholinyl-N-oxide, 1-oxo-1-thiomorpholinyl, pyridinonyl, benzimidazolonyl, benzo[d]oxazol-2(3H)-onyl, 3,4-dihydroisoquinolin-1(2H)-onyl, indolin-onyl, 1H-imidazo[4,5-c]pyridin-2(3H)-onyl, 7H-purin-8(9H)-onyl, imidazolidin-2-onyl, 1H-imidazol-2(3H)-onyl, 1,1-dioxo-1-thiomorpholinyl, and the like.
• oxo ( ⁇ O) or oxide (—O—) substituents such as piperid
• Disease refers to any disease, disorder, condition, symptom, or indication.
• Halo or “halogen” refers to a fluoro, chloro, bromo, or iodo group.
• Haloalkyl refers to an alkyl group in which at least one hydrogen is replaced with a halogen.
• haloalkyl includes monohaloalkyl (alkyl substituted with one halogen atom) and polyhaloalkyl (alkyl substituted with two or more halogen atoms).
• Representative “haloalkyl” groups include difluoromethyl, 2,2,2-trifluoroethyl, 2,2,2-trichloroethyl, and the like.
• perhaloalkyl means, unless otherwise stated, an alkyl group in which each of the hydrogen atoms is replaced with a halogen atom.
• perhaloalkyl includes, but is not limited to, trifluoromethyl, pentachloroethyl, 1,1,1-trifluoro-2-bromo-2-chloroethyl, and the like.
• Heteroaryl refers to a monovalent heteroaromatic group derived by the removal of one hydrogen atom from a single atom of a parent heteroaromatic ring system. Heteroaryl groups typically include 5- to 14-membered, but more typically include 5- to 10-membered aromatic, monocyclic, bicyclic, and tricyclic rings containing one or more, for example, 1, 2, 3, or 4, or in certain embodiments, 1, 2, or 3, heteroatoms chosen from O, S, or N, with the remaining ring atoms being carbon. In monocyclic heteroaryl groups, the single ring is aromatic and includes at least one heteroatom.
• a monocyclic heteroaryl group may include 5 or 6 ring members and may include 1, 2, 3, or 4 heteroatoms, 1, 2, or 3 heteroatoms, 1 or 2 heteroatoms, or 1 heteroatom where the heteroatom(s) are independently selected from O, S, or N.
• bicyclic aromatic rings both rings are aromatic.
• bicyclic heteroaryl groups at least one of the rings must include a heteroatom, but it is not necessary that both rings include a heteroatom although it is permitted for them to do so.
• heteroaryl includes a 5- to 7-membered heteroaromatic ring fused to a carbocyclic aromatic ring or fused to another heteroaromatic ring.
• the rings are aromatic and at least one of the rings includes at least one heteroatom.
• the point of attachment may be at the ring including at least one heteroatom or at a carbocyclic ring.
• the total number of S and O atoms in the heteroaryl group exceeds 1, those heteroatoms are not adjacent to one another.
• the total number of S and O atoms in the heteroaryl group is not more than 2.
• the total number of S and O atoms in the aromatic heterocycle is not more than 1.
• Heteroaryl does not encompass or overlap with aryl as defined above.
• heteroaryl groups include, but are not limited to, groups derived from acridine, carbazole, cinnoline, furan, imidazole, indazole, indole, indolizine, isobenzofuran, isochromene, isoindole, isoquinoline, isothiazole, 2H-benzo[d][1,2,3]triazole, isoxazole, naphthyridine, oxadiazole, oxazole, perimidine, phenanthridine, phenanthroline, phenazine, phthalazine, pteridine, purine, pyrazine, pyrazole, pyridazine, pyridine, pyrimidine, pyrrole, pyrrolizine, quinazoline, quinoline, quinolizine, quinoxaline, tetrazole, thiadiazole, thiazole, thiophene, triazo
• the heteroaryl group can be between 5 to 20 membered heteroaryl, such as, for example, a 5 to 14 membered or 5 to 10 membered heteroaryl.
• heteroaryl groups can be those derived from thiophene, pyrrole, benzothiophene, 2H-benzo[d][1,2,3]triazole benzofuran, indole, pyridine, quinoline, imidazole, benzimidazole, oxazole, tetrazole, and pyrazine.
• MTAP refers to a mammalian methylthioadenosine phosphorylase enzyme.
• “Pharmaceutically acceptable” refers to generally recognized for use in animals, and more particularly in humans.
• “Pharmaceutically acceptable salt” refers to a salt of a compound that is pharmaceutically acceptable and that possesses the desired pharmacological activity of the parent compound.
• “Pharmaceutically acceptable excipient” refers to a broad range of ingredients that may be combined with a compound or salt of the present invention to prepare a pharmaceutical composition or formulation.
• excipients include, but are not limited to, diluents, colorants, vehicles, anti-adherents, glidants, disintegrants, flavoring agents, coatings, binders, sweeteners, lubricants, sorbents, preservatives, and the like.
• PRMT5 refers to a mammalian Protein Arginine N-Methyl Transferase 5 (PRMT5) enzyme.
• PRMT5 inhibitor refers to compounds that inhibit or negatively modulate all or a portion of the PRMT5 enzymatic activity.
• MTA-cooperative PRMT5 inhibitor refers to compounds that inhibit or negatively modulate all or a portion of the PRMT5 enzymatic activity in the presence of bound MTA, in vitro or in vivo, in the cells with elevated levels of MTA.
• Stepoisomer refers to an isomer that differs in the arrangement of the constituent atoms in space. Stereoisomers that are mirror images of each other and optically active are termed “enantiomers,” and stereoisomers that are not mirror images of one another and are optically active are termed “diastereomers.”
• Subject includes mammals and humans.
• the terms “human” and “subject” are used interchangeably herein.
• “Therapeutically effective amount” refers to the amount of a compound that, when administered to a subject for treating a disease, or at least one of the clinical symptoms of a disease or disorder, is sufficient to affect such treatment for the disease, disorder, or symptom. As those skilled in the art will recognize. this amount is typically not limited to a single dose but may comprise multiple dosages over a significant period of time as required to bring about a therapeutic or prophylactic response in the subject. Thus, a “therapeutically effective amount” is not limited to the amount in a single capsule or tablet, but may include more than one capsule or tablet, which is the dose prescribed by a qualified physician or medical care provider.
• the “therapeutically effective amount” can vary depending on the compound, the disease, disorder, and/or symptoms of the disease or disorder, severity of the disease, disorder, and/or symptoms of the disease or disorder, the age of the subject to be treated, and/or the weight of the subject to be treated. An appropriate amount in any given instance can be readily apparent to those skilled in the art or capable of determination by routine experimentation.
• Treating” or “treatment” of any disease or disorder refers to arresting or ameliorating a disease, disorder, or at least one of the clinical symptoms of a disease or disorder, reducing the risk of acquiring a disease, disorder, or at least one of the clinical symptoms of a disease or disorder, reducing the development of a disease, disorder or at least one of the clinical symptoms of the disease or disorder, or reducing the risk of developing a disease or disorder or at least one of the clinical symptoms of a disease or disorder.
• Treating” or “treatment” also refers to inhibiting the disease or disorder, either physically, (e.g., stabilization of a discernible symptom), physiologically, (e.g., stabilization of a physical parameter), or both, or inhibiting at least one physical parameter which may not be discernible to the subject. Further, “treating” or “treatment” refers to delaying the onset of the disease or disorder or at least symptoms thereof in a subject which may be exposed to or predisposed to a disease or disorder even though that subject does not yet experience or display symptoms of the disease or disorder.
• compositions that include the compound or the pharmaceutically acceptable salt thereof, the tautomer thereof, the pharmaceutically acceptable salt of the tautomer, the stereoisomer of any of the foregoing, or the mixture thereof according to any one of the examples and at least one pharmaceutically acceptable excipient, carrier or diluent.
• the compound or the pharmaceutically acceptable salt thereof, the tautomer thereof, the pharmaceutically acceptable salt of the tautomer, the stereoisomer of any of the foregoing, or the mixture thereof according to any one of the aspects is present in an amount effective for the treatment of PRMT5-dependent cancers.
• the pharmaceutical composition is formulated for oral delivery whereas in other embodiments, the pharmaceutical composition is formulated for intravenous delivery.
• the pharmaceutical composition is formulated for oral administration once a day or QD, and in some such formulations is a tablet where the effective amount of the active ingredient ranges from 1 mg to 1000 mg.
• the subject is a mammal. In some such aspects, the mammal is a rodent. In other aspects, the mammal is a canine. In still other embodiments, the subject is a primate and, in some embodiments, is a human.
• compositions or formulations for the administration of the compounds of this invention may conveniently be presented in unit dosage form and may be prepared by any of the methods well known in the art. All methods include the step of bringing the active ingredient into association with the carrier which constitutes one or more accessory ingredients.
• the pharmaceutical compositions are prepared by uniformly and intimately bringing the active ingredient into association with a liquid carrier or a finely divided solid carrier or both, and then, if necessary, shaping the product into the desired formulation.
• the active object compound is included in an amount sufficient to produce the desired effect upon the process or condition of diseases.
• the compounds of the invention may be administered via oral, mucosal (including sublingual, buccal, rectal, nasal, or vaginal), parenteral (including subcutaneous, intramuscular, bolus injection, intra-arterial, or intravenous), transdermal, or topical administration.
• the compounds of the invention are administered via mucosal (including sublingual, buccal, rectal, nasal, or vaginal), parenteral (including subcutaneous, intramuscular, bolus injection, intra-arterial, or intravenous), transdermal, or topical administration.
• the compounds of the invention are administered via oral administration.
• the compounds of the invention may find use in treating a number of conditions.
• compositions described herein are generally useful for the inhibition of PRMT5.
• methods of treating PRMT5-mediated disorder in a subject comprise administering an effective amount of a compound described herein (e.g., a compound of Formula I or a pharmaceutically acceptable salt thereof), to a subject in need of treatment.
• the effective amount is a therapeutically effective amount.
• the effective amount is a prophylactically effective amount.
• the subject is suffering from a PRMT5-mediated disorder (e.g., a cancer, for example a lymphoma, breast cancer, or pancreatic cancer).
• the subject is susceptible to a PRMT5-mediated disorder (e.g., a cancer, for example a lymphoma, breast cancer, or pancreatic cancer).
• PRMT5-mediated disorder means any disease, disorder, or other pathological condition in which PRMT5 is known to play a role. Accordingly, in some aspects, the present disclosure relates to treating or lessening the severity of one or more diseases in which PRMT5 is known to play a role.
• herein provided is a method of inhibiting PRMT5 activity in a subject in need thereof comprising administering to the subject an effective amount of a compound described herein (e.g., a compound of Formula I, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof.
• a compound described herein e.g., a compound of Formula I, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof.
• the invention provides methods of treating cancers and other disorders arising from homozygous deletion of the chromosome 9p21 locus, which harbors the well-known tumor suppressor CDKN2A (cyclin dependent kinase inhibitor 2A).
• the invention encompasses methods of treating cancers and tumors which are MTAP (methylthioadenosine phosphorylase)-null.
• these types of cancer display accumulation of MTAP substrate, methylthioadenosine (MTA).
• the methods of treating PRMT5 disorders encompassed by the invention preferentially target PRMT5 in MTAP null tumors while sparing PRMT5 in normal tissues (MTAP WT).
• the compounds of the present invention thus include MTA-cooperative small molecule inhibitors which could preferentially target the MTA bound state of PRMT5, enriched in MTAP null tumor cells, while providing an improved therapeutic index over normal cells where MTAP is intact and MTA levels are low.
• a PRMT5 inhibitor MTA cooperative compound contemplated by the present invention is useful in treating a proliferative disorder, such as cancer.
• the cancer compounds described herein are useful for treating pancreatic cancer.
• the cancer compounds described herein are useful for treating multiple myeloma (MM).
• the cancer compounds described herein are useful for treating breast cancer.
• the breast cancer can be estrogen receptor negative (ER ⁇ ) or the breast cancer can be progesterone receptor negative (PR ⁇ ).
• the breast cancer can be HER2 negative.
• the breast cancer is estrogen receptor negative, progesterone receptor negative and HER2 negative, also referred to herein as “triple negative breast cancer”.
• a breast cancer can be a lobular carcinoma in situ (LCIS), a ductal carcinoma in situ (DCIS), an invasive ductal carcinoma (IDC), inflammatory breast cancer, Paget disease of the nipple, Phyllodes tumor, Angiosarcoma, adenoid cystic carcinoma, low-grade adenosquamous carcinoma, medullary carcinoma, mucinous carcinoma, papillary carcinoma, tubular carcinoma, metaplastic carcinoma, micropapary carcinoma, mixed carcinoma, or another breast cancer, including but not limited to triple negative, HER positive, estrogen receptor positive, progesterone receptor positive, HER and estrogen receptor positive, HER and progesterone receptor positive, estrogen and progesterone receptor positive, and HER and estrogen and progesterone receptor positive.
• compounds of the invention are useful for treating pancreatic cancer.
• compounds of the invention are useful for treating NSCLC (non-small cell lung carcinoma.
• NSCLC non-small cell lung carcinoma.
• the NSCLC can be squamous NSCLC. In another embodiment, it can be adenocarcinoma.
• cancer can be glioblastoma (GBM). In a further aspect, cancer can be mesothelioma. In one aspect, cancer can be bladder cancer. In another aspect, cancer can be esophageal cancer. In a further aspect, cancer can be melanoma. In one aspect, cancer can be DLBCL, HNSCC or cholangiocarcinoma.
• GBM glioblastoma
• cancer can be mesothelioma.
• cancer can be bladder cancer.
• cancer can be esophageal cancer.
• cancer can be melanoma.
• cancer can be DLBCL, HNSCC or cholangiocarcinoma.
• one or more compounds described herein are useful for treating any PRMT5-mediated or PRMT5-responsive proliferative cell disorder, for example a cancer that is PRMT5 responsive.
• a cancer that lacks p53 is less sensitive to PRMT5 inhibition than a cancer that is p53 positive.
• a cancer that is PRMT5 responsive can be a p53 positive cancer.
• the term “p53 positive” refers to a cancer that does not lack p53 expression and/or activity.
• one or more compounds described herein are useful for treating a p53 positive cancer.
• a greater amount of one or more compounds described herein may be required to treat a p53 negative cancer (e.g., a p53 null cancer) than a p53 positive cancer.
• the disclosure provides a method for identifying subjects having a cancer that is sensitive to treatment with a PRMT5 inhibitor.
• the method comprises obtaining a sample from the subject; detecting the presence or absence of p53; and, identifying the subject as having a cancer that is sensitive to treatment with a PRMT5 inhibitor if p53 is present in the sample.
• a subject having a p53 positive cancer is identified as a subject for treatment with a PRMT5 inhibitor.
• the method further comprises administering to the subject a composition comprising a PRMT5 inhibitor.
• aspects of the disclosure relate to a method for identifying subjects having a cancer that is insensitive (or that has low sensitivity) to treatment with a PRMT5 inhibitor.
• the method comprises obtaining a sample from the subject; detecting the presence or absence of p53; and, identifying the subject as having a cancer that is not sensitive (for example, a cancer that is less sensitive than a p53 positive cancer) to treatment with a PRMT5 inhibitor if p53 is absent from the sample (e.g., if the cancer is a p53 null cancer).
• a p53 negative cancer (e.g., a p53 null cancer) is treated with a PRMT5 inhibitor, but a greater amount of PRMT5 inhibitor may be required to treat the p53 negative cancer than a p53 positive cancer.
• a subject having a p53 negative cancer (e.g., a p53 null cancer) is treated with a therapeutic agent that is not a PRMT5 inhibitor.
• sample any biological sample derived from the subject, includes but is not limited to, cells, tissues samples, body fluids (including, but not limited to, mucus, blood, plasma, serum, urine, saliva, and semen), cancer cells, and cancer tissues.
• Detection of the presence or absence of p53 in the sample may be achieved by any suitable method for detecting p53 nucleic acid or protein, for example, nucleic acid sequencing (e.g., DNA or RNA sequencing), quantitative PCR, Western blotting, etc., or any combination of thereof.
• one or more of the compounds described herein may be useful for treating other types of cancer, including, but not limited to, acoustic neuroma, adenocarcinoma, adrenal gland cancer, anal cancer, angiosarcoma (e.g., lymphangiosarcoma, lymphangioendotheliosarcoma, hemangio sarcoma), appendix cancer, benign monoclonal gammopathy, biliary cancer (e.g., cholangiocarcinoma), bladder cancer, brain cancer (e.g., meningioma; glioma, e.g, astrocytoma, oligodendroglioma; medulloblastoma), bronchus cancer, carcinoid tumor, cervical cancer (e.g., cervical adenocarcinoma), choriocarcinoma, chordoma, craniopharyngioma, colorectal cancer (e.
• angiosarcoma
• HCC hepatocellular cancer
• lung cancer e.g., bronchogenic carcinoma, small cell lung cancer (SCLC), non-small cell lung cancer (NSCLC), adenocarcinoma of the lung
• myelofibrosis MF
• chronic idiopathic myelofibrosis chronic myelocytic leukemia (CML), chronic neutrophilic leukemia (CNL), hypereosinophilic syndrome (HES)
• neuroblastoma e.g., neurofibromatosis (NF) type 1 or type 2, schwannomatosis
• neuroendocrine cancer e.g., gastroenteropancreatic neuroendoctrine tumor (GEP-NET), carcinoid tumor
• osteosarcoma ovarian cancer (e.g., cystadenocarcinoma, ovarian embryonal carcinoma, ovarian adenocarcinoma), papillary adenocarcinoma
• penile cancer e.g., Paget's disease of the penis and scrotum
• pinealoma e.g., primitive neuroectodermal tumor (PNT)
• prostate cancer e.g., prostate adenocar
• the method of treating cancer in a subject comprises administering a composition comprising a PRMT5 inhibitor to the subject, wherein treatment with the PRMT5 inhibitor inhibits tumor growth of the cancer by more than about 25%, more than about 50%, more than about 75%, more than about 90% (e.g., 25%-50%, 50%-75%, 75%-90%, or 90%-100% for example).
• the method of treating cancer in a subject comprises administering a composition comprising a PRMT5 inhibitor to the subject, wherein methyl mark of the cancer is reduced more than about 50%, more than about 75%, more than about 80% (e.g., 50%-75%, 50%-80%, 80%-90%, 80%-100%, or 90%-100% for example).
• a methyl mark refers to protein methylation, for example a histone methylation (e.g., methylation of one or more lysines and/or arginines of a histone protein), or DNA methylation (e.g., epigenetic DNA methylation, for example methylated CpG sites).
• the methyl mark level of a cell is a measure of the extent to which histones are methylated in the cell (e.g., at one or more particular lysine and/or arginine positions).
• Method A Compound I can be prepared from the reaction of acid IA and secondary amine IB-1 in the presence of a base such as Et 3 N or DIPEA, an activating reagent such as HATU or PyBrOP, in a solvent such as DMF or DMAc. If racemic amine or acid is employed in Method A, chiral SFC can be used to separate the stereoisomers, in which case stereochemistry was arbitrarily assigned to each isomer.
• Method B Compound I can be prepared from the reaction of acid chloride IC and secondary amine IB in the presence of a base such as Et 3 N or DIPEA or pyridine, in a solvent such as THF or dioxane or DCM or DCE. Alternatively, compound I can be prepared from the reaction of acid chloride IC and secondary amine IB in the presence of DMAP in pyridine. If racemic amine or acid is employed in Method B, chiral SFC can be used to separate the stereoisomers, in which case stereochemistry was arbitrarily assigned to each isomer.
• Step 1 A vial was charged with 3-chloro-6-(difluoromethoxy)pyridazine (675 mg, 3.74 mmol), sodium carbonate (1190 mg, 11.2 mmol), and [1,1′-bis(diphenylphosphino)ferrocene] dichloropalladium (II) (274 mg, 0.374 mmol). 1,4-dioxane (16.8 mL) and water (1.87 mL) were then added, followed by vinylboronic acid pinacol ester (1.73 g, 1.90 mL, 11.2 mmol). The resulting mixture was sparged with nitrogen for 15 min and then heated to 80° C. After 15 h, the mixture was cooled to 23° C.
• Step 2 A vial was charged with 3-(difluoromethoxy)-6-vinylpyridazine (573 mg, 3.33 mmol), acetone (7.93 mL), and water (1.59 mL). To the resulting solution was added potassium osmate (VI) dihydrate (123 mg, 0.333 mmol) followed by 4-methylmorpholine 4-oxide (1.37 g, 11.7 mmol), and the resulting mixture was allowed to stir at 23° C.
• Step 3 A vial was charged with 1-(6-(difluoromethoxy)pyridazin-3-yl)ethane-1,2-diol (300 mg, 1.45 mmol) and tetrahydrofuran (13.4 mL). To the resulting solution were added sodium (meta)periodate (933 mg, 4.36 mmol) and water (1.12 mL), and the resulting mixture was allowed to stir at 23° C. After 1 h, the reaction mixture was diluted with H 2 O (10 mL), transferred to a separatory funnel with CH 2 Cl 2 (20 mL) and brine (20 mL), and extracted with CH 2 Cl 2 (2 ⁇ 20 mL).
• Step 1 A round-bottom flask was charged with sodium hydride (60% dispersion in mineral oil, 1.556 g, 38.9 mmol) and tetrahydrofuran (31.1 mL). The headspace was flushed with nitrogen, and the mixture was cooled to 0° C. Subsequently, cyclopropanol (2.281 g, 1.901 mL, 39.3 mmol) was added, and the resulting mixture was allowed to warm to 23° C. under nitrogen for 1 h. 3-chloro-6-methylpyridazine (1.00 g, 7.78 mmol) was then added dropwise as a solution in tetrahydrofuran (8.0 mL), and the reaction mixture was allowed to stir at 23° C.
• sodium hydride 50% dispersion in mineral oil, 1.556 g, 38.9 mmol
• tetrahydrofuran 31.1 mL
• Step 2 A vial was charged with 3-cyclopropoxy-6-methylpyridazine (308.5 mg, 2.054 mmol), selenium dioxide (365 mg, 3.29 mmol), and 1,4-dioxane (8.22 mL). The resulting mixture was sparged with nitrogen for 10 min, and the vial was subsequently heated to 110° C. After 30 min, the reaction mixture was allowed to cool to 23° C. and was filtered over a pad of Celite (30 mL 3:1 EtOAc:EtOH eluent) and concentrated to dryness.
• a vial was charged with 3-ethoxy-6-methylpyridazine (1.07 g, 7.72 mmol), selenium dioxide (1.37 g, 12.3 mmol), and 1,4-dioxane (30.9 mL). The resulting mixture was sparged with nitrogen for 10 min, and the vial was subsequently heated to 110° C. After 30 min, the reaction mixture was allowed to cool to 23° C. and was filtered over a 1 cm pad of Celite (30 mL 3:1 EtOAc:EtOH eluent) and conc. to dryness.
• Step 1 An oven-dried vial was charged with 4 ⁇ Molecular Sieves (2.00 g, 2.64 mmol), dichloromethane (10.6 mL), and 6-methoxypyridazine-3-carbaldehyde (365 mg, 2.64 mmol, Enamine). To the resulting suspension was added 1-((tributylstannyl)methoxy)propan-2-amine (SnAP 3Me-M reagent, 0.907 mL, 2.64 mmol) via syringe, and the resulting mixture was allowed to stir at 23° C.
• SnAP 3Me-M reagent 1-((tributylstannyl)methoxy)propan-2-amine
• Step 2 A vial was charged oven-dried copper (II) trifluoromethanesulfonate (191 mg, 0.528 mmol), (4S,4'S)-2,2′-(propane-2,2-diyl)bis(4-phenyl-4,5-dihydrooxazole) (88 mg, 0.264 mmol), (4R,4′R)-2,2′-(propane-2,2-diyl)bis(4-phenyl-4,5-dihydrooxazole) (88 mg, 0.264 mmol), and hexafluoroisopropanol (4 mL). The resulting mixture was stirred at 23° C. for 7 h.
• Step 3 The racemic secondary amine 4 (337 mg) was purified via preparative SFC using a Chiral Technologies IG column (250 ⁇ 21 mm, 5 mm) with a mobile phase of 50% liquid CO 2 and 50% MeOH with 0.2% TEA using a flowrate of 80 mL/min. Peak assignment determined by SFC with IG column with 50% MeOH with 0.2% TEA.
• the 1st eluting peak was arbitrarily assigned as (3S,5R)-3-(6-methoxypyridazin-3-yl)-5-methylmorpholine (5, 150.7 mg, >99% ee) and the 2 nd eluting peak was arbitrarily assigned as (3R,5S)-3-(6-methoxypyridazin-3-yl)-5-methylmorpholine (6, 152.7 mg, >99% ee).
• Racemic amines summarized in Table 1 were prepared in a fashion similar to that described above for amine 4.
• Step 1 As performed in Angew. Chem. Int. Ed. 2016, 55, 9676-9: to a vigorously stirred mixture in a 25-mL reaction vial of Boc-1-homoPro(4,4-difluoro) (1.050 g, 3.96 mmol, RSP Amino Acids, LLC), 4,5,6,7-tetrachloro-2-hydroxyisoindoline-1,3-dione (1.191 g, 3.96 mmol, Aldrich) and 4-(dimethylamino) pyridine (0.048 g, 0.396 mmol, Sigma-Aldrich Corporation) in DCM (25 mL) was dropwise added at rt N,N′-Diisopropylcarbodiimide (0.550 g, 0.674 mL, 4.35 mmol, Sigma-Aldrich Corporation) via a syringe.
• Boc-1-homoPro(4,4-difluoro) 1.050 g,
• Step 2 A 25-mL reaction vessel charged with nickel (ii) chloride hexahydrate (50.3 mg, 0.212 mmol, Sigma-Aldrich Corporation) and 1,10-bathophenanthroline (70.3 mg, 0.212 mmol, Combi-Blocks Inc.) was subjected to evacuation followed by back-filling with argon (3 ⁇ ) before N, N-dimethylformamide (4.10 mL) was introduced. The resulting mixture was stirred under argon at rt for 2.5 h as a green solution.
• Step 3 To a stirred solution of tert-butyl 2-(4-bromophenyl)-4,4-difluoropiperidine-1-carboxylate (290 mg, 0.771 mmol, impure from Step 2) in DCM (5 mL) was added at rt 2,2,2-trifluoroacetic acid (88 mg, 4.0 mL, 0.771 mmol, Aldrich). The resulting mixture was stirred at rt for 1 h.
• Racemic amines in Table 3 were prepared in a fashion similar to that described above for amine 57.
• Step 1 To a 150-mL round-bottomed flask was added methyl 4-amino-3-bromobenzoate (4 g, 17.39 mmol, Combi-Blocks Inc.) and bis(pinacolato)diboron (8.83 g, 34.8 mmol, Frontier Scientific, Inc.) in 1,4-dioxane (58.0 mL). To the solution was then added potassium acetate (5.12 g, 52.2 mmol, Sigma-Aldrich Corporation), the mixture was degassed by bubbling through with Argon for 5 minutes.
• Step 2 To a stirred solution of 4-oxotetrahydrofuran-3-carbonitrile (0.500 g, 4.50 mmol) in dichloromethane (5.00 mL) was added DIPEA (0.943 mL, 5.40 mmol) and the reaction mixture was cooled to ⁇ 78° C. Then, triflic anhydride (0.760 mL, 4.50 mmol) was added dropwise at ⁇ 78° C. for 1 min and the reaction mixture stirred at same temperature for 15 min.
• reaction mixture was diluted with water, the organic layer was separated, washed with brine (2 ⁇ 10 mL), dried over sodium sulfate, and concentrated to give crude 4-cyano-2,5-dihydrofuran-3-yl trifluoromethanesulfonate (1.05 g, 4.32 mmol, 96% yield), which was used in the next step without further purification.
• Step 3 To a stirred solution of 4-cyano-2,5-dihydrofuran-3-yl trifluoromethanesulfonate (10 g, 41.1 mmol) in 1,4-dioxane (200 mL) and water (20.00 mL) was added methyl 4-amino-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzoate (9.12 g, 32.9 mmol), K 2 CO 3 (17.05 g, 123 mmol), and Pd(PPh 3 ) 4 (4.75 g, 4.11 mmol) under nitrogen purging. Then, the reaction mixture heated at 80° C. for 16 h.
• Step 4 To a stirred solution of methyl 4-amino-1,3-dihydrofuro[3,4-c]quinoline-8-carboxylate (30 g, 123 mmol) in water (300 mL):tetrahydrofuran (300 mL):methanol (300 mL) was added LiOH (11.77 g, 491 mmol) and the reaction mixture heated at 75° C. for 3 h. The reaction mixture concentrated and then the aqueous layer acidified with 1.5 N HCl up to pH 6.0.
• Step 1 A mixture of methyl 2-oxocyclopentanecarboxylate (1.0 g, 0.877 mL, 7.03 mmol, Matrix Scientific) and 1,1′-dimethyltriethylamine (1.000 g, 1.352 mL, 7.74 mmol, Sigma-Aldrich Corporation) in DCM (15 mL) was cooled to ⁇ 78° C. and trifluoromethanesulfonic acid anhydride (7.03 mL, 7.03 mmol, Sigma-Aldrich Corporation) was added. After complete addition, the mixture was stirred at ⁇ 78° C. for 5 min, then the dry ice-bath was removed and stirred at rt.
• Step 2 A mixture of methyl 2-(((trifluoromethyl)sulfonyl)oxy)cyclopent-1-ene-1-carboxylate (1.982 g, 7.23 mmol), (2-amino-5-(methoxycarbonyl)pyridin-3-yl)boronic acid (1.70 g, 8.67 mmol), potassium phosphate, tribasic (3.78 g, 21.69 mmol, Acros) and [1,1′-bis(diphenylphosphino)ferrocene]dichloropalladium(ii), complex with dichloromethane (0.177 g, 0.217 mmol, Strem Chemicals, Inc.) in 1,4-dioxane/water (10/0.60 mL) was heated at 80° C.
• Step 3 A mixture of methyl 6-oxo-6,7,8,9-tetrahydro-5H-cyclopenta[c][1,8]naphthyridine-2-carboxylate (1.76 g, 7.21 mmol) in POCl 3 (24.68 g, 15 mL, 161 mmol, Aldrich) was heated to reflux for 30 min. The reaction went to completion and was carefully added to cold-sat. aq NaHCO 3 to basify the reaction.
• Step 4 To a suspension of methyl 6-chloro-8,9-dihydro-7H-cyclopenta[c][1,8]naphthyridine-2-carboxylate (1.89 g, 7.19 mmol) in DMSO (15 mL) was added DIPEA (2.79 g, 3.77 mL, 21.58 mmol, Aldrich) followed by the addition of (2,4-dimethoxyphenyl)methanamine (1.564 g, 1.405 mL, 9.35 mmol, Aldrich). The resulting mixture was heated at 90° C. overnight. The reaction was cooled to rt, diluted with water, washed with sat. NH 4 Cl and extracted with EtOAc.
• Step 5 To a solution of methyl 6-((2,4-dimethoxybenzyl)amino)-8,9-dihydro-7H-cyclopenta[c][1,8]naphthyridine-2-carboxylate (2.18 g, 5.54 mmol) in THF/MeOH (10/10 mL) was added NaOH (10 mL, 10.00 mmol) and the resulting solution was heated at 70° C. for 2 h. When the reaction reached completion, it was brought to rt and acidified with 10 mL 1M HCl.
• Step 1 A mixture of 3-furancarboxylic acid, tetrahydro-4-oxo methyl ester (3.0 g, 3.00 mL, 20.82 mmol, Ambeed, Inc.) and DIPEA (2.96 g, 4.00 mL, 22.90 mmol, Aldrich) in DCM (20 mL) was cooled to ⁇ 78° C. and trifluoromethanesulfonic anhydride (20.82 mL, 20.82 mmol, Sigma-Aldrich Corporation) was added. After complete addition, the mixture was stirred at ⁇ 78° C. for 5 min then the dry ice-bath was removed and stirred at rt. After 15 min the LCMS showed desired mass.
• Step 2 A mixture of methyl 4-(((trifluoromethyl)sulfonyl)oxy)-2,5-dihydrofuran-3-carboxylate (2.349 g, 8.50 mmol), (5-amino-2-(methoxycarbonyl)pyridin-4-yl)boronic acid (2.0 g, 10.21 mmol), potassium phosphate, tribasic (4.44 g, 25.5 mmol, Acros) and [1,1′-bis(diphenylphosphino)ferrocene]dichloropalladium (ii), complex with dichloromethane (0.347 g, 0.425 mmol, Strem Chemicals, Inc.) in 1,4-dioxane/water (20/1.20 mL) was heated at 90° C.
• Step 3 A mixture of methyl 4-oxo-1,3,4,5-tetrahydrofuro[3,4-c][1,7]naphthyridine-8-carboxylate (2.0 g, 8.12 mmol) and POCl 3 (32.9 g, 20 mL, 215 mmol, Aldrich) was heated to reflux for 3 h. The reaction was brought to rt and carefully added to cold-sat. NaHCO 3 to basify the reaction. After stirring for 15 min, the mixture was extracted with EtOAc and the combined organics were concentrated to afford methyl 4-chloro-1,3-dihydrofuro[3,4-c][1,7]naphthyridine-8-carboxylate. m/z (ESI): 265 (M+H) + . Theoretical yield was considered.
• Step 4 To a mixture methyl 4-chloro-1,3-dihydrofuro[3,4-c][1,7]naphthyridine-8-carboxylate (2.15 g, 8.12 mmol) in DMSO (20 mL) was added DIPEA (3.15 g, 4.26 mL, 24.37 mmol, Aldrich) followed by the addition of (2,4-dimethoxyphenyl)methanamine (1.630 g, 1.464 mL, 9.75 mmol, Aldrich). The resulting mixture was heated at 90° C. overnight. The reaction was brought to rt, diluted with water and extracted with EtOAc.
• Step 5 To a solution of methyl 4-((2,4-dimethoxybenzyl)amino)-1,3-dihydrofuro[3,4-c][1,7]naphthyridine-8-carboxylate (0.500 g, 1.264 mmol) in THF/MeOH (10/10 mL) was added NaOH (5.0 mL, 5.00 mmol, EDM) and the resulting solution was heated at 70° C. for 2 h. Then, the reaction was brought to rt and acidified with 1M HCl (5 mL).
• Step 1 To a suspension of sodium hydride (11.10 g, 278 mmol 0.5 equiv., 60% in mineral oil) in anhydrous tetrahydrofuran (250 mL) was added methyl 2-hydroxyacetate (42.4 mL, 555 mmol, 1.0 equiv) at room temperature under N 2 atmosphere. To the reaction mixture (E)-but-2-enenitrile (54.5 mL, 666 mmol, 1.0 equiv) was added slowly at 65° C. and stirred for 2 h at same temperature. The reaction mixture was cooled and quenched with 2N NaOH solution (250 mL) and extracted with diethyl ether (500 mL). The aqueous layer was acidified with conc.
• Step 2 To a stirred solution of 2-methyl-4-oxotetrahydrofuran-3-carbonitrile (25.0 g, 200 mmol, 1.0 equiv) in dichloromethane (500 mL) was added DIPEA (69.8 mL, 400 mmol, 2.0 equiv) and triflic anhydride (47.1 mL, 280 mmol, 1.4 equiv) at ⁇ 78° C. and stirred at same temperature for 15 min. The reaction mixture was quenched with slow addition of water (250 mL) and after attaining the room temperature was extracted with dichloromethane (2 ⁇ 500 mL). The combined organic layer was dried over sodium sulfate, filtered and concentrated under reduced pressure.
• DIPEA 69.8 mL, 400 mmol, 2.0 equiv
• triflic anhydride 47.1 mL, 280 mmol, 1.4 equiv
• Step 3 To a stirred solution of 4-cyano-5-methyl-2,5-dihydrofuran-3-yl trifluoromethanesulfonate (35 g, 136 mmol, 1.0 equiv) in 1,4-dioxane (1400 mL) and water (70.0 mL), was added methyl 4-amino-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzoate (37.7 g, 136 mmol, 1.0 equiv) and potassium phosphate tribasic (87 g, 408 mmol, 3.0 equiv) under nitrogen atmosphere.
• reaction mixture was degassed with nitrogen for 15 min and then PdCl 2 (dppf)-DCM adduct (9.96 g, 13.61 mmol, 0.1 equiv) was added and the reaction mixture was heated at 90° C. for 16 h.
• LCMS indicated completion of the reaction.
• the reaction mass was concentrated under reduced pressure to get crude product.
• the crude residue was purified by column chromatography over silica gel (60-120 mesh) using 50% ethyl acetate with hexanes as an eluent to give methyl 4-amino-3-methyl-1,3-dihydrofuro[3,4-c]quinoline-8-carboxylate (25 g, 97 mmol, 71% yield) as a brown solid.
• Step 4 To a stirred solution of methyl 4-amino-3-methyl-1,3-dihydrofuro[3,4-c]quinoline-8-carboxylate (26.0 g, 101 mmol, 1.0 equiv) in tetrahydrofuran (130 mL), methanol (78 mL) and water (52 mL), was added lithium hydroxide (9.64 g, 403 mmol, 4.0 equiv) and stirred at 75° C. for 4 h. LCMS indicated completion of the reaction. The reaction mixture was concentrated under reduced pressure. The crude residue was dissolved in water (100 mL) and filtered to removed insoluble particles. The aqueous layer was acidified with con.
• Step 5 Chiral SFC separation: 44.5 g of racemic 4-amino-3-methyl-1,3-dihydrofuro[3,4-c]quinoline-8-carboxylic acid was separated by chiral SFC to get 14 g of each isomer. Stereochemistry is assigned arbitrarily.
• Step 1 To a stirred solution of diethyl (cyanomethyl)phosphonate (130 g, 732 mmol, 1.1 equiv) in tetrahydrofuran (2000 mL) was added potassium tert-butoxide (1M solution in THF; 732 mL, 732 mmol, 1.1 equiv) at ⁇ 78° C. and stirred for 30 min. To the reaction mixture, 2-(benzyloxy)acetaldehyde (100 g, 666 mmol, 1.0 equiv) was added at ⁇ 78° C. and allowed the mixture to warm to rt over 1 h.
• 2-(benzyloxy)acetaldehyde 100 g, 666 mmol, 1.0 equiv
• Step 2 To a stirred solution of potassium tert-butoxide (1M solution in THF; 289.0 mL, 289 mmol, 1.0 equiv) in tetrahydrofuran (260 mL) was added methyl 2-hydroxyacetate (22.03 mL, 289 mmol, 1.0 equiv) at RT and heated to 50° C. under nitrogen atmosphere. To this, 4-(benzyloxy)but-2-enenitrile (50.0 g, 289 mmol, 1.0 equiv) was added and stirred at same temperature for 4 h. Reaction monitored by TLC. Reaction temperature was increased up to 70° C. and stirred for 16 h. After completion, the reaction mixture was cooled to 0° C.
• Step 3 To a stirred solution of 2-((benzyloxy)methyl)-4-oxotetrahydrofuran-3-carbonitrile (5.8 g, 25.08 mmol, 1.0 equiv) in dichloromethane (116 mL) were added Triflic anhydride (6.75 mL, 40.1 mmol, 1.6 equiv) and DIPEA (8.76 mL, 50.2 mmol, 2.0 equiv) at ⁇ 78° C. under N 2 atmosphere and stirred for 10 min. The reaction mixture was quenched with water (50 mL) and extracted with dichloromethane (2 ⁇ 200 mL).
• Step 4 To a stirred solution of 5-((benzyloxy)methyl)-4-cyano-2,5-dihydrofuran-3-yl trifluoromethane sulfonate (7.65 g, 20.93 mmol, 1.0 equiv) in 1,4-dioxane (232 mL) and water (11.60 mL) were added methyl 4-amino-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzoate (5.8 g, 20.93 mmol, 1.0 equiv), potassium carbonate (8.68 g, 62.8 mmol, 3.0 equiv) at room temperature.
• Reaction mixture was purged with N 2 gas for 15 min and then added Pd(PPh 3 ) 4 (1.209 g, 1.046 mmol, 0.05 equiv).
• the reaction mixture was heated at 80° C. for 16 h. After completion, the reaction mixture was concentrated under reduced pressure and the crude residue was purified by column chromatography over silica gel (60-120 mesh) using 80% ethyl acetate with pet ether as eluent to give 4-amino-3-((benzyloxy)methyl)-1,3-dihydrofuro[3,4-c]quinoline-8-carboxylate (4.4 g, 58% yield) as an off white solid.
• Ester 151 was treated with LiOH in THF (similar to Step 4 to intermediate 146) and the lithium salt of 151 was used crude in amide coupling reactions.
• Step 1 To a stirred suspension of methyl 3-fluoro-4-nitrobenzoate (1.07 g, 5.37 mmol, 1.00 equiv) and 1H-imidazole-2-carbonitrile (0.500 g, 5.37 mmol, 1.0 equiv) in dimethyl sulfoxide (5.00 mL) was added DIPEA (2.35 mL, 13.43 mmol, 2.5 equiv.) at room temperature and the reaction mixture was stirred 16 hours. The reaction mixture was then concentrated under reduced pressure to obtain crude material that was diluted with EtOAc and washed with water. The layers were separated and organic layer was dried over anhydrous Na 2 SO 4 and concentrated under reduced pressure.
• Step 2 To a stirred solution of methyl 3-(2-cyano-1H-imidazol-1-yl)-4-nitrobenzoate (0.900 g, 3.31 mmol, 1.0 equiv) in tetrahydrofuran (10.0 mL) and water (2.00 mL) was added acetic acid (0.946 mL, 16.53 mmol, 5.0 equiv.) and iron powder (1.85 g, 33.1 mmol, 10.0 equiv) at room temperature. The reaction mixture was stirred for 16 hours. The reaction mixture was then diluted with EtOAc and washed with satd. aq NaHCO 3 solution.
• Step 1 To the solution of ethyl 5-methyl-1H-pyrazole-4-carboxylate (5.00 g, 32.4 mmol, 1.0 equiv, Combi-Blocks) in acetic acid (100 mL) was added bromine (5.01 mL, 97.0 mmol, 3.0 equiv) and sodium acetate (10.6 g, 130 mmol, 4.0 equiv.) at rt. Then the reaction mixture was stirred and heated for 16 h. The reaction was slowly quenched with sodium bicarbonate and extracted with ethyl acetate.
• Step 2 To a stirred solution of ethyl 3-bromo-5-methyl-1H-pyrazole-4-carboxylate (4.80 g, 20.6 mmol, 1.0 equiv) in dichloromethane (15 mL) was added DHP (2.26 mL, 24.7 mmol, 1.2 equiv) and tosic acid (0.78 g, 4.12 mmol, 0.2 equiv) at 0° C. The resulting reaction mixture was stirred for 16 h to completion. The reaction was quenched with water (20 mL) and extracted with ethyl acetate (20 mL ⁇ 3).
• Step 3 To a stirred solution of methyl 4-amino-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzoate (7.34 g, 26.5 mmol, 1.2 equiv, LabNetwork) in 1,4-dioxane (112 mL) and water (28.0 mL) was added ethyl 3-bromo-5-methyl-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazole-4-carboxylate (7.00 g, 22.1 mmol, 1.0 equiv), potassium phosphate, tribasic (9.36 g, 44.1 mmol, 2.0 equiv) under nitrogen purging for 10 min at room temperature.
• Step 4 To a stirred solution of methyl 4-hydroxy-3-methyl-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazolo[4,3-c]quinoline-8-carboxylate (600 mg, 1.76 mmol, 1.0 equiv) in dichloromethane (3.00 mL). Then trifluoromethanesulfonic anhydride (992 mg, 3.52 mmol, 2.0 equiv) and DIPEA (921 ⁇ L, 5.27 mmol, 3.0 equiv) was added and the reaction mixture kept between 30-32° C. for 16 h.
• reaction mixture was concentrated under reduced pressure to get 300 mg (31% crude yield) of crude methyl 3-methyl-1-(tetrahydro-2H-pyran-2-yl)-4-(((trifluoromethyl)sulfonyl)oxy)-1H-pyrazolo[4,3-c]quinoline-8-carboxylate.
• Step 5 A solution of methyl 4-((4-methoxybenzyl)amino)-3-methyl-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazolo[4,3-c]quinoline-8-carboxylate (2.80 g, 6.08 mmol, 1.0 equiv) in trifluoroacetic acid (28.0 mL) was heated at 90° C. for 12 h. The reaction mixture was concentrated under reduced pressure to get crude methyl 4-amino-3-methyl-1H-pyrazolo[4,3-c]quinoline-8-carboxylate (3.50 g, 13.7 mmol, 225% crude yield). m/z: 257.3 [M+H] + .
• Step 6 To a stirred solution of methyl 4-amino-3-methyl-1H-pyrazolo[4,3-c]quinoline-8-carboxylate (3.50 g, 13.7 mmol, 1.0 equiv) in tetrahydrofuran (35.0 mL), methanol (35.0 mL), water (35.0 mL) at room temperature. Then lithium hydroxide monohydrate (4.02 g, 96.0 mmol, 7.0 equiv) was added and the reaction mixture was stirred at rt for 16 h. The reaction mixture was quenched with water and a solid precipitate was observed. The solid was filtered and dried under vacuum.
• Step 1 K 3 PO 4 H 2 O (1.08 g, 4.70 mmol, Sigma-Aldrich Corporation), X-Phos (0.08 g, 0.16 mmol, Sigma-Aldrich Corporation), (2-dicyclohexylphosphino-2′,4′,6′-triisopropyl-1,1′-biphenyl)[2-(2′-amino-1,1′-biphenyl)]palladium (ii) methanesulfonate (0.14 mg, 0.16 mmol, Sigma-Aldrich Corporation), 1-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1 h-pyrazole-4-carbonitrile (1.10 g, 4.70 mmol, Enamine) and methyl 4-amino-5-bromo-2-(trifluoromethyl)benzoate (0.700 g, 2.349 mmol, Combi Blocks) were suspended in a degas
• Step 2 Methyl 4-amino-1-methyl-7-(trifluoromethyl)-1H-pyrazolo[4,3-c]quinoline-8-carboxylate (0.62 g, 1.90 mmol) and lithium hydroxide (0.91 g, 3.79 mmol, Sigma-Aldrich Corporation) were suspended in methanol (3.0 mL), H 2 O (3.0 mL) and THF (3.0 mL) and stirred at 50° C. for 2 hours.
• Example 7 Examples in Table 7 were prepared in a manner similar to that described above for Example 300 using the indicated amide coupling reagent in the table.
• Examples 452 and 453 ((R)-4-amino-3-methyl-1,3-dihydrofuro[3,4-c][1,7]naphthyridin-8-yl)((S)-3-(4-(trifluoromethyl)phenyl)morpholino)methanone and ((S)-4-amino-3-methyl-1,3-dihydrofuro[3,4-c][1,7]naphthyridin-8-yl)((S)-3-(4-(trifluoromethyl)phenyl)morpholino)methanone
• 100 mg of the racemate product was purified via preparative SFC using a Chiral Technologies OJ column (250 ⁇ 21 mm, 5 mm) with a mobile phase of 75% liquid CO 2 and 25% MeOH with 0.2% TEA using a flowrate of 80 mL/min. to generate 39.7 mg of peak 1 with an ee of >99% and 40.5 mg of peak 2 with an ee of 99.8%. Peak assignment determined by SFC with OJ column with 25% MeOH with 0.2% TEA. Stereochemistry is assigned arbitrarily.
• Examples in Table 8 were prepared in a manner similar to that described above for example 452 and 453 using the indicated amide coupling reagent in the table and purification conditions.
• Step 1 To a solution of 3-(4-(trifluoromethyl)phenyl)morpholine (0.100 g, 0.432 mmol, Enamine), 4-((2,4-dimethoxybenzyl)amino)-1,3-dihydrofuro[3,4-c][1,7]naphthyridine-8-carboxylic acid hydrochloride (138) (0.271 g, 0.649 mmol) and 1,1′-dimethyltriethylamine (0.559 g, 0.755 mL, 4.32 mmol, Sigma-Aldrich Corporation) in DMF (4 mL) was added bromotripyrrolidinophosphonium hexafluorophosphate (0.202 g, 0.432 mmol, Sigma-Aldrich Corporation) and the resulting mixture was heated at 50° C.
• Step 2 (S)-(4-amino-1,3-dihydrofuro[3,4-c][1,7]naphthyridin-8-yl)(3-(4-(trifluoromethyl)phenyl)morpholino)methanone and (R)-(4-amino-1,3-dihydrofuro[3,4-c][1,7]naphthyridin-8-yl)(3-(4-(trifluoromethyl)phenyl)morpholino)methanone (4-amino-1,3-dihydrofuro[3,4-c][1,7]naphthyridin-8-yl)(3-(4-(trifluoromethyl)phenyl)morpholino)methanone 2,2,2-trifluoroacetate were separated via preparative SFC using a Chiral Technologies AD column (150 ⁇ 21 mm, 5 mm) with a mobile phase of 60% Liquid CO 2 and 40% MeOH with 0.2%
• Peak 1 (S)-(4-amino-1,3-dihydrofuro[3,4-c][1,7]naphthyridin-8-yl)(3-(4-(trifluoromethyl)phenyl)morpholino)methanone (481) as a white solid.
• Peak 2 (R)-(4-amino-1,3-dihydrofuro[3,4-c][1,7]naphthyridin-8-yl)(3-(4-(trifluoromethyl)phenyl)morpholino)methanone (482) as a white solid.
• Example 483 rac-(4-amino-1,3-dihydrofuro[3,4-c][1,8]naphthyridin-8-yl)(3-(4-(trifluoromethyl)phenyl)morpholino)methanone
• Example 483 was prepared in a manner similar to compound 481-rac above. m/z (ESI): 445.1 [M+H] +
• Example 484 was prepared in a manner similar to compound 481-rac above using commercial enantiopure (3)-3-[4-(Trifluoromethoxy)phenyl]morpholine hydrochloride (NetChem, CAS #1391448-60-4). m/z (ESI): 474.0 [M+H] +
• Example 481 and 482 were prepared in a manner similar to that described for Example 481 and 482.
• Step 1 To a stirred suspension of 4-amino-1,3-dihydrofuro[3,4-c][1,7]naphthyridine-8-carboxylic acid (500 mg, 2.163 mmol) in CH 2 Cl 2 (10.0 mL) was added 4 M HCl in 1,4-dioxane (1.62 mL, 6.49 mmol) and the resulting suspension was allowed to stir at room temperature for 30 min. The mixture was concentrated under reduced pressure, then co-evaporated with toluene (2 ⁇ 5 mL).
• the obtained crude material was re-suspended in dichloromethane (20.0 mL), cooled to 0° C., and treated with oxalyl chloride (2.0 M in CH 2 Cl 2 , 4.33 mL, 8.65 mmol) followed by DMF (5 drops).
• the reaction vessel was flushed with nitrogen and the reaction mixture was allowed to stir at room temperature under nitrogen overnight.
• Step 2 A vial was charged with (3S,5R)-3-(6-methoxypyridazin-3-yl)-5-methylmorpholine (35.9 mg, 0.172 mmol), 4-amino-1,3-dihydrofuro[3,4-c][1,7]naphthyridine-8-carbonyl chloride hydrochloride (63.8 mg, 0.223 mmol) and dichloromethane (3.43 mL). To the resulting suspension was added N,N-diisopropylethylamine (111 mg, 150 ⁇ L, 0.858 mmol), and the resulting mixture was stirred at rt for 16 h. The reaction was quenched by addition of sat. aq.
• the sample was purified by SFC using column Chiralpak IC (250 ⁇ 21 mm, 5 ⁇ m), with a modifier of 25% MeOH+ TEA using a flowrate of 100 mL/min.
• Submitted sample was 70 mg dissolved in 10 mL MeOH:DCM 1:1 (7 mg/mL).
• Example 11 Examples in Table 11 were prepared in a manner similar to that described for Example 566 and 567.
• 603 (4-amino-1,3-dihydrofuro[3,4- c]quinolin-8-yl)((3S)-3-(4- (2,2,2-trifluoroethyl)phenyl)- 4-morpholinyl)methanone 458 1 st peak, Chiralcel AS- H column (250 ⁇ 21 mm, 5 um) with a mobile phase of 80% Liquid CO 2 and 20% MeOH + TEA using a flowrate of 80 mL/min 604 (4-amino-1,3-dihydrofuro[3,4- c][1,7]naphthyridin-8- yl)((3S)-3-(4-(2,2,2- trifluoroethyl)phenyl)-4- morpholinyl)methanone 459 1 st peak, Chiral Technologies AS column (250 ⁇ 21 mm, 5 mm) with a mobile phase of 80% Liquid CO 2 and 20% MeOH with 0.2%
• Example 626 was prepared in an identical manner to example 625. m/z (ESI): 461.10 [M+H] +
• an HCT-116 isogenic cell line pair was utilized where one cell line was engineered to genetically knockout both MTAP alleles. Cell viability was then assessed in both the parent HCT-116 cell line and the MTAP null cell line after 6 days of treatment with compounds of the present invention. Selective anti-proliferative activity in the MTAP null cell line indicates MTA-cooperative inhibition of PRMT5 and ability to inhibit growth of cancer cells that have loss of MTAP.
• HCT116 MTAP null and WT cells were seeded in 96-well tissue culture plates in RPMI 1640 media+10% fetal bovine serum. Plates were incubated overnight at 37° C. and 5% CO 2 . Cells were then treated with an 8- or 9-point serial dilution of compound, using a top concentration of 1, or 10 ⁇ M, 1:3 serial dilution steps and, a DMSO-only control. Cells were incubated in the presence of drug for 6 days. Effects on cell viability were measured with the CellTiter-Glo® Luminescent Cell Viability Assay (Promega) per manufacturer's recommendation.
• IC 50 values were calculated with GraphPad Prism v 5.01 using symmetrical sigmoidal dose-response least squares fit with Hill slope fixed to ⁇ 1 and top constrain to 100% or GeneData Screener using a 4-parameter logistic model to fit dose response curves.
• HCT116-MTAP null and WT cell line proliferation HCT-116 HCT-116 MTAP null WT IC 50

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